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Macronutrients Content in Leaves of Red Raspberry ‘Willamette’ as
Affected by Foliar Fertilization and Substrate
ić, Đ. Ružić, T. Milošević
M. Pešaković and R. Miletić Facult
Fruit Research Institute
Keywords: tissue culture, fertilization, macroelements imbalance
Raspberry (Rubus idaeus L.) cultivar ‘Willamette’ has long been most com-
monly grown raspberry cultivar in Serbia, which is owing to high adaptability of the
cultivar to respective agro-environmental conditions. Massive dieback of full bearing
plantings is a major problem in raspberry growing hence quality planting material is
a must when establishing new raspberry plantings. This study was conducted under
protected conditions (in screenhouse) on plants obtained by micropropagation in vitro.
Over a two-year period plants were grown on two substrates (Steckmedium, Seedling)
and treated with three foliar fertilizers (Wuxal, Murtonik and Ferticare). In terms of
macroelements content, the studies suggested nitrogen excess (3.09–3.41%), but also
deficiency in phosphorus (0.14-0.20%), potassium (1.25-1.32%), calcium (1.03-1.07%)
and magnesium (0.26-0.30%). The assessment of nutritional status of plants by DOP
index suggested significant differences in microelements imbalance when different
foliar fertilizers and substrates are applied.
From the economic aspect, red raspberry (Rubus idaeus L.) is a major soft fruit in
Serbia (Central Serbia), the total production of fresh fruits amounting to 58,000 tons
(FAOSTAT, 2011). According to Nikolić et al. (2008), cultivar ‘Willamette’ predominates
in the Serbian raspberry growing, accounting for 95% of the total area.
In recent years, a massive dieback of raspberry plants has been observed in
raspberry plantings of Serbia, which affects the exploitation period. A species of
Phytophthora spp. genus are believed to be the primary causal agent of the phenomenon
(Koprivica et al., 2002). However, the fact that raspberry plantings were established with
shoots originating from commercial plantings considerably contributed to the dieback.
In order to ensure steady and quality yields, quality planting material is a must for
planting establishment. Healthy, true-to-type plants are obtained by micropropagation in
vitro (Ružić and Lazić, 2004), and optimal and balanced plant nutrition ensures high plant
vegetative potential. Ružić et al. (2005) report on a long utilization of foliar fertilizers in
fruit growing, and issues raised pertaining to this method of plant nutrition are mainly
concerned with the efficacy of this nutrition method. Only when nutrients are firmly fixed
in the soil by means of particular chemical processes, and when requirements of some
plant cultures for particular nutrients are higher than the adsorption, foliar nutrition is a
necessary agro-technical measure (Neilsen and Neilsen, 1986). This nutrition mode is not
particularly effective in satisfying plants’ needs for macroelements, compared to their
requirements for microelements, due to the already absorbed nutrients in the plant (Finck,
1985; Kirkby and Roemheld, 2004). Generally, relatively little is known about essential
processes in the plant, such as nutrient uptake in leaves, nutrients processing in the plant
and their effect on different physiological processes in leaves and fruits (Schlegel and
The objective of this investigation was to study indirect impact of applied foliar
nutrition and different substrates on macroelements content in raspberry leaves which
meet the criteria in terms of health and true-to-typeness.
Balkan Symposium on Fruit Growing
Eds.: M. Coman and E. Chitu
Acta Hort. 981, ISHS 2013
MATERIALS AND METHODS
Plants of raspberry ‘Willamette’ obtained by micropropagation in vitro were used
for planting establishment. The research was conducted in screenhouse (30x6 m) on a site
of Fruit Research Institute, Čačak. The screenhouse is protected by anti-insect net
ensuring full plant isolation and protection. The study involved three foliar fertilizers
[Wuxal–0.3%, Murtonik–0.2%, Ferticare (Kemira–0.5%) and no treatment in the control
variant] and two plant growth substrates (Steckmedium, Seedling). Given the three foliar
fertilizers and the control, the trial involved four variants of treatment (80 plants) in four
replications with 20 plants in each experimental unit. Some 640 plants were potted on
12th May 2004 in plastic 0.5 dm3 vessels and transferred into 2 dm3 plastic pots in 2005.
Both substrates are slightly acid (pH=6), enriched with water-soluble nutrients,
microelements and wetting agents. The following foliar fertilizers were applied: Wuxal
(N–8%; P2O5–8%; K2O–6%; B–0.01%; Mn–0.013%; Cu–0.007%; Fe–0.0015%; Mo–
0.001%; Zn–0.005%); Murtonik (N–19%; P2O5–9%; K2O–27% and microelements) and
Ferticare-Kemira (N–24%; P2O5–8%; K2O–16%; S–5.0%; Mg–3.7%; Fe–0.1%; Mn–
0.05%; B–0.03%; Cu–0.01%; Zn–0.025%; Mo–0.004%). Plants were treated periodically,
i.e., every fifteen days. First treatment in 2004 was done on 1st June (two weeks after
planting), and the last on 15th July. In 2005, first treatment was done on 10th June, and the
last on 25th July.
Raspberry leaf samples, used to determine macroelements content, were collected
from all the evaluated treatments (late July, 2005). Leaves were prepared for chemical
analysis, which involved rinsing in a solution of nitric acid, air drying, and drying at 65°C
(in a drier). Nitrogen content was assessed by Kjeldahl’s method (Tecator-Kjeltec System
1003; Hoganas, Sweden). Potassium, calcium and manganese contents were determined
by the atomic absorption spectrophotometer (SP 191-Pye Unicam, England), whereas
phosphorus, assessed with spectrophotometer (PU 8740 UV/VIS, England) and expressed
in % of leaf dry matter. The DOP index of leaf macroelements was calculated for each
substrate and foliar fertilizer by:
where: C stands for nutrient content in a sample studied, and Cref is nutrient
content considered as optimum, both values being given on a dry matter basis. The Cref
was taken from macroelements optimum values, as proposed by Kessel (2003). The
ΣDOP is obtained by adding the DOP index values irrespective of sign. The deviation
from optimum percentage (DOP index) was estimated for the diagnosis of mineral content
status in leaves (Montañés et al., 1991). The larger the ΣDOP, the greater the nutrients
imbalance was (Zarrouk et al., 2005).
Experimental data were subjected to analysis of variance (ANOVA) using
MSTAT-C statistical computer package (Michigan State University, East Lansing, MI,
USA). Dunnett’s test (d') at d'≤ 0.05 and d' ≤ 0.01 was used for mean separation of
experimental data referring to the application of foliar fertilizers. The Duncan’s Multiple
Range Test was used for mean separation of data concerning application of substrates as
well as for analysis of interaction among substrates and foliar fertilizers. Testing of ΣDOP
mean values was based on LSD test at p ≤ 0.05.
RESULTS AND DISCUSSION
Table 1 shows the results pertaining to macroelements content in leaves of rasp-
berry plants (‘Willamette’) grown on different substrates and treated with different
fertilizers. Analysis of variance of macroelements content suggests the existence not only
of significant differences in P and K content resulting from application of different
substrates, but also in P and Mg content in plants treated with foliar fertilizers. Addi-
tionally, substrate/foliar fertilizer interaction also had a marked influence on N and P
Phosphorus content in raspberry leaves ranged between 0.14±0.00% and
0.21±0.02%. Considerably higher macroelements content was found in leaves of plants
developing on Steckmedium (0.17±0.01%). Application of different foliar fertilizers
reveals higher P content, compared to other variants of treatment, in leaves of plants
treated with Murtonik (0.21±0.02%), which is the only significant aspect of difference
compared to the control.
Spiers (2002) reports on similar results whereby application of varying formula-
tions of different foliar fertilizers (0 Mg/l, control, 10control, 10N) in blackberry
resulted in higher P content in leaves. According to Warman (2009), application of
adequate substrates for growing raspberry ‘Willamette’ contributes to the N and P uptake.
The results presented in Table 1 indicate that K content in leaves of studied
raspberry plants ranged from 1.25±0.01% to 1.32±0.02%. Considerably higher K content
was recorded in leaves of plants developing on Steckmedium (1.32±0.02%). Plant growth
substrates for plants transferred from tissue culture laboratory to greenhouse or in open
field for plant acclimatization usually need to be composed of sterile earth, sterilized
sand, or their mixtures and vermiculite (Mišić, 1998).
Application of different foliar fertilizers gave significant differences in Mg
content. It was highest in plants treated with Ferticare (0.30±0.01%), and the lowest in
those sprayed with Wuxal, as well as in the control (0.30±0.01%). Significant differences
in Mg content compared to the control were evidenced only in case of treating plants with
Figures 1 and 2 show considerable influence of interaction effect of substrate/
foliar fertilizer on N and P content.
The results pertaining to macroelements content in raspberry leaves based on DOP
index are presented in Table 2. The analysis of results pointed to N surplus and P, K, Ca
and Mg deficiencies as the result of utilization of the different growing substrates.
According to Lučić et al. (1995), excess nitrogen may be due to the high mobility of
nitrogen, i.e., its easy movement towards intensively growing organs and tissues up to
their development-safe limit. Given the fact that leaf samples were collected during mid-
growing period (end of July), and that according to Kastori (1986) the highest phosphorus
content in leaves is at the early stages of growing period, P deficiency is quite expected.
According to Ubavić et al. (2001), insufficient Ca and Mg supply is due to their poor
mobility from the place of uptake.
Analysis of variance suggests major contribution of interaction between the
substrate and foliar fertilizer to macronutrients imbalance (Table 2). The maximum
deviation from optimal balance of macroelements was observed in plants grown on
Seedling (ΣDOP = 155.19), whereas the lowest was in those treated with Murtonik
(ΣDOP = 125.95). DOP index values and ΣDOP provide similar information to the
Diagnosis and Recommendation Integrated System (DRIS) (Davee et al., 1986; Sanz,
1999; Jimenez, 2007). The deviation from optimal balance by using different substrates
and foliar fertilizers can be accounted for by a hindered establishment of a balanced
nutrition in fruit trees compared to other agricultural crops (Veličković, 2006).
The analysis of mineral composition of leaves of raspberry ’Willamette’ points to
the excess N and P, K, Ca and Mg deficiencies.
The highest deviation from optimal macroelements balance was observed in plants
grown on Seedling, while it was lowest in plants treated with foliar fertilizer Murtonik.
This study is the part of the project No. 31093 financed by Ministry of Education,
Science and Technological Development of the Republic of Serbia. We hereby express
our sincere gratitude for the support.
Davee, D.E., Righetti, T.L., Fallahi, E. and Robbins, S. 1986. An evaluation of the DRIS
approach for identifying mineral limitations on yield in ‘Napolean’ sweet cherry.
Journal of the American Society for Horticultural Science 111:988-993.
Finck, A. 1985. Fertilizer and Fertilization. p.179-236. Weinheim-Deerfield Florida,
Food and Agricultural Organization. 2011. www.fao.org .
Jiménez, S., Pinochet, J., Gogorcena, Y., Betrán, J.A. and Moreno, M.A. 2007. Influence
of different vigour cherry rootstocks on leaves and shoots mineral composition.
Scientia Horticulturae 112(1):73-79.
Kastori, R. 1986. Fiziologija biljaka II. Biblioteka matice Srpske, Novi Sad.
Kessel, C. 2003. Fertilizing raspberries – raspberry leaf analysis. Fruit Production Recom-
mendations. Publication 360:1-3.
Kirkby, E.A. and Roemheld, V. 2004. Micronutrients in plant physiology: functions,
uptake and mobility. Proceedings of the International Fertiliser Society, York, 543:1-52.
Koprivica, M., Milenković, S., Gavrilović, V. and Milijašević, S. 2002. Nove bolesti
maline i manje poznate interakcije štetočina i patogena maline. XII simpozijum o
zaštiti bilja i primeni pesticida, Zlatibor (Srbija). 56 p.
Lučić, P., Đurić, G. and Mićić, N. 1995. Voćarstvo I. Partenon, Institut Srbija, Beograd.
Mišić, P.D. 1998. Malina. Zajednica za voće i povrće d.d., Beograd.
Montañés, L., Heras, L. and Sanz, M. 1991. Desviacién del óptimo porcentual (DOP):
nuevo indice para la interpretación del análisis vegetal. An Aula Dei 20:93-107.
Neilsen, G.H. and Neilsen, D. 1986. Tree fruit zinc nutrition. p.85-93. In: A.B. Petersen
and R.G. Stevens (eds.), Good Fruit Grower, Chap. 10, Yakima.
Nikolić, M., Ivanović, M., Milenković, S., Milivojević, J. and Milutinović, M. 2008. The
state and prospects of raspberry production in Serbia. Acta Hort. 777:243-250.
Ružić, Đ. and Lazić, T. 2004. Mikropropagacija maline cv Willamette in vitro. Jugo-
slovensko voćarstvo 145-146:109-117.
Ružić, Đ., Ličina, V., Stikić, R., Cerović, R., Vulić, T. and Ruml, M. 2005. New investiga-
tion tendencies in physiology and ecology of fruits. Journal of Pomology 39(152):
Sanz, M. 1999. Evaluation of interpretation of DRIS system during growing season of the
peach tree: comparison with DOP method. Soil Science and Plant Analysis 30:1025-
Schlegel, T.K. and Schönher, J. 2002. Penetration of calcium chloride into apple fruits as
affected by stage of fruit development. Acta Hort. 594:527-533.
Spiers, J.M. 2002. Influence of N, P, K, Ca and Mg rate on leaf macronutrient concen-
tration of ‘Navaho’ blackberry. Acta Hort. 585:659-663.
Ubavić, M., Kastori, R., Oljača, R. and Marković, M. 2001. Ishrana voćaka. Poljoprivredni
fakultet Univerziteta u Banjaluci i Naučno voćarsko društvo Republike Srpske,
Veličković, M. 2006. Voćarstvo. Poljoprivredni fakultet, Beograd.
Warman, P.R. 2009. Soil and plant response to applications of municipal solid waste
compost and fertilizer to Willamette raspberries. International Journal of Fruit Science
Zarrouk, O., Gogorcena, Y., Gómez-Aparisi, J., Betrán, J.A. and Moreno, M.A. 2005.
Influence of almond × peach hybrids rootstock on flower and leaf mineral content,
yield and vigour of two peach cultivars. Scientia Horticulturae 106:502-514.
Table 1. Macroelements content in leaves of raspberry ‘Willamette’ in the second year
after planting (2005).
Treatment Macroelements content (%)
N P K Ca Mg
Seedling 3.29±0.13a 0.14±0.00b 1.25±0.01b 1.06±0.02a 0.28±0.01a
Steckmedium 3.21±0.72a 0.17±0.01a 1.32±0.02a 1.05±0.02a 0.29±0.00a
Wuxal 3.10±0.11ns 0.20±0.01ns 1.29±0.04ns 1.05±0.03ns 0.27±0.01ns
Murtonik 3.09±0.12ns 0.21±0.02** 1.29±0.03ns 1.04±0.03ns 0.29±0.01ns
Ferticare 3.40±0.18ns 0.20±0.01ns 1.26±0.02ns 1.07±0.03ns 0.30±0.01**
Control 1 3.41±0.16 0.20±0.01 1.30±0.02ns 1.05±0.03 0.27±0.01
A ns ** * ns ns
B ns ** ns ns *
A B ** ** ns ns ns
1 The control is a plant not treated with a foliar fertilizer.
‘A’ and ‘B’ stand for substrate and foliar fertilizer respectively.
Repeated small letter is not significantly different at (P≤0.05) by Duncan’s Multiple Range test.
* indicate significant difference between means of the control and other foliar fertilizers at d’≤ 0.05 and d’
≤ 0.01 by Dunnett’s test.
ns – non significant difference.
Table 2. Macroelements content in ‘Willamette’ raspberry plants in the second year after
planting (2005) based on DOP index.
Treatment Macroelements ∑ DOP
N P K Ca Mg
Substrate Seedling +19.64 -60.00 -16.66 -34.57 -24.32 155.19 a
Steckmedium +16.73 -51.43 -12.00 -36.36 -21.62 138.14 b
Wuxal +12.73 -42.86 -14.00 -36.36 -27.03 132.98 b
Murtonik +12.36 -40.00 -14.00 -36.97 -21.62 125.95 a
Ferticare +23.64 -42.86 -16.00 -35.15 -18.92 136.57 c
Control1 +24.00 -42.86 -13.33 -36.36 -27.03 143.58 d
1 The control is a plant not treated with a foliar fertilizer.
(-) indicates lower macroelements supply as compared to the optimal content.
(+) indicates higher macroelements supply as compared to the optimal content.
Values within each column followed by the same small letter are not significantly different at p≤0.05 by
Fig. 1. Nitrogen content (AB). Fig. 2. Phosphorus content (AB).
* Repeated small letter is not significantly different at (P≤0.05) by Duncan’s multiple range test.
Wuxal Murtonik Ferticare Control
Wuxal Murtonik Ferticare Control